Identification of triosephosphate isomerase (TIM) genes from Microcystis (Cyanobacteria : Chroococcales) and theoretical analyses of the potential of cyanobacterial TIM as a target for designing specific inhibitors

The genes encoding triosephosphate isomerase (TIM) in three species of Microcystis (M. aeruginosa, M. viridis and M. wesenbergii) were investigated. Reverse transcriptase-polymerase chain reaction indicated that they were transcribed in the cells. Analyses showed that their DNA and deduced amino acid sequences were highly conserved between all the three species, only a single nonsynonymous substitution was seen at position 31, from an Asp in M. aeruginosa and M. viridis to Glu in M. wesenbergii. Sequence alignment of these with 12 other known cyanobacterial TIM sequences showed that all the cyanobacterial TIMs had a very high level of amino acid identity (over 50% between each two). Comparison of the cyanobacterial TIMs with other reported TIMs (from diverse lineages of the three Domains) showed that they possessed common active-site residues and sequence motifs. All cyanobacterial TIMs have two common cysteine residues (Cys127 and Cys176), and the Cys176 is almost cyanobacteria-specific with only one exception in Streptomyces coelicolor. Both secondary structure alignment and comparative modelling of Synechocystis sp. TIM showed that Cys176 was located at the hinge region of the flexible loop-6 and might therefore be critical to the movement of TIM's loop-6, which is important to the function of the enzyme. Thus, the cyanobacterial TIM-specific Cys176 may be a potential site for the discovery of suitable drugs against cyanobacteria, and such drugs may have utility in controlling water blooms due to cyanobacteria.

Diversity Variation of Ground-dwelling Beetle(Insecta:Coleoptera) Communities in Jiaozi Mountain, Yunnan, China

对云南轿子雪山自然保护区地表甲虫物的种组成及生物多样性季节变化进行了连续跟踪调查.调查结果如下:(1)通过对4种典型微环境样地(草地、灌丛、林地以及农田生态系统)连续3次跟踪连续调查,共获得标本2451头,分别隶属于24科.其中步甲科(Carabidae)为优势类群,占总数的62.10%;隐翅虫科(Staphylinidae)其次,占12.77%.可能由于海拔、气温等因素影响,该地区昆虫活动高峰期较短.(2)通过对4种典型微生态环境中地表甲虫的生物多样性的比较,表明不同生境内甲虫的多样性指数存在动态变化,在不同生境片区内甲虫存在迁移活动,甲虫多在灌丛中越冬,并随季节及食物源迁移.

Consumption of fa cai Nostoc soup: A Potential for BMAA exposure from Nostoc cyanobacteria in China?

Grown in arid regions of western China the cyanobacterium Nostoc flagelliforme - called fa cai in Mandarin and fat choy in Cantonese - is wild-harvested and used to make soup consumed during New Year's celebrations. High prices, up to $125 USD/kg, led to overharvesting in Inner Mongolia, Ningxia, Gansu, Qinghai, and Xinjiang. Degradation of arid ecosystems, desertification, and conflicts between Nostoc harvesters and Mongol herdsman concerned the Chinese environmental authorities, leading to a government ban of Nostoc commerce. This ban stimulated increased marketing of a substitute made from starch. We analysed samples purchased throughout China as well as in Chinese markets in the United States and the United Kingdom. Some were counterfeits consisting of dyed starch noodles. A few samples from California contained Nostoc flagelliforme but were adulterated with starch noodles. Other samples, including those from the United Kingdom, consisted of pure Nostoc flagelliforme. A recent survey of markets in Cheng Du showed no real Nostoc flagelliforme to be marketed. Real and artificial fa cai differ in the presence of beta-N-methylamino-L-alanine (BMAA). Given its status as a high-priced luxury food, the government ban on collection and marketing, and the replacement of real fa cai with starch substitutes consumed only on special occasions, it is anticipated that dietary exposure to BMAA from fa cai will be reduced in the future in China.

PCR-DGGE Fingerprinting Analysis of Plankton Communities and Its Relationship to Lake Trophic Status

Plankton communities in eight lakes of different trophic status near Yangtze, China were characterized by using denatured gradient gel electrophoresis (DGGE). Various water quality parameters were also measured at each collection site. Following extraction of DNA from plankton communities, 16S rRNA and 18S rRNA genes were amplified with specific primers for prokaryotes and eukaryotes, respectively; DNA profiles were developed by DGGE. The plankton community of each lake had its own distinct DNA profile. The total number of bands identified at 34 sampling stations ranged from 37 to 111. Both prokaryotes and eukaryotes displayed complex fingerprints composed of a large number of bands: 16 to 59 bands were obtained with the prokaryotic primer set; 21 to 52 bands for the eukaryotic primer set. The DGGE-patterns were analyzed in relation to water quality parameters by canonical correspondence analysis (CCA). Temperature, pH, alkalinity, and the concentration of COD, TP and TN were strongly correlated with the DGGE patterns. The parameters that demonstrated a strong correlation to the DGGE fingerprints of the plankton community differed among lakes, suggesting that differences in the DGGE fingerprints were due mainly to lake trophic status. Results of the present study suggest that PCR-DGGE fingerprinting is an effective and precise method of identifying changes to plankton community composition, and therefore could be a useful ecological tool for monitoring the response of aquatic ecosystems to environmental perturbations.

Characteristics of the microbial communities in the integrated vertical-flow constructed wetlands

Microorganisms play an important role in removing pollutants from constructed wetlands. We investigated the microbial characteristics in a novel integrated vertical-flow constructed wetland (IVCW), which has been in operation in Wuhan, China since 1998. We used phospholipid fatty acid (PLFA) and amoA gene to analyze the structure and diversity of the microbial community within the IVCW. PLFA results suggested that the amount of bacterial PLFA was significantly higher than that of fungal PLFA, but the total microbial biomass represented by PLFA index was low in the system. Microbial spatial distribution showed significantly higher bacterial (both G(+) and G(-)) and fungal biomass in the surface than in the subsurface layers. The ratios of monounsaturated to branched PLFA demonstrated that an anaerobic layer sandwiched by two aerobic layers existed in the IVCW, consistent with the redox potential results. Analysis of the amoA revealed the presence of Nitrosomonas-like sequences in the surface substrate of the downflow chamber and apparent diversities of ammonia-oxidizing bacteria in the system. These results suggest that microorganisms, despite their relatively low biomass, have inhabited the IVCW, and the results will offer some valuable information on microbe to system designers and managers.

Effects of dietary cyanobacteria of two different sources on growth and recovery of hybrid tilapia (Oreochromis niloticus x O. aureus)

A 115 days feeding trial was conducted to evaluate the effect of dietary cyanobacteria on growth, microcystins (MCs) accumulation in hybrid tilapia (Oreochromis niloticus x O. aureus) and the recovery when the fish were free of cyanobacteria. Three experimental diets were formulated: the control (cyanobacteria free diet); one test diet with cyanobacteria from Lake Taihu (AMt 80.0 mu g MCs g(-1) diet) and one with cyanobacteria from Lake Dianchi (AMd, 410.0 rho g MCs g(-1) diet). Each diet was fed to fish for 60 days and then all fish were free of cyanobacteria for another 55 days. A significant increase in feeding rate (FR) was observed in fish fed AMd diet after a first 30-day exposure (1(st) EP), and in fish fed both AMt diet and AMd diet after a second 30-day exposure (2(nd) EP). Specific growth rates (SGR) of fish fed AMt diet and AMd diet were both obviously affected after the first 30-day exposure, but SGR was only significantly affected in fish fed AMt diet after the second 30-day exposure. After a 55-day recovery, there were no significant differences among diets in the indices mentioned above. Much higher concentrations of MCs were accumulated in tissues of all fish exposed to cyanobacteria. After the 55-day recovery, MC concentrations in fish tissues were significantly lower than those on day 60. (C) 2009 Elsevier Ltd. All rights reserved.

Communities of gastrointestinal helminths of fish in historically connected habitats: habitat fragmentation effect in a carnivorous catfish Pelteobagrus fulvidraco from seven lakes in flood plain of the Yangtze River, China

Background: Habitat fragmentation may result in the reduction of diversity of parasite communities by affecting population size and dispersal pattern of species. In the flood plain of the Yangtze River in China, many lakes, which were once connected with the river, have become isolated since the 1950s from the river by the construction of dams and sluices, with many larger lakes subdivided into smaller ones by road embankments. These artificial barriers have inevitably obstructed the migration of fish between the river and lakes and also among lakes. In this study, the gastrointestinal helminth communities were investigated in a carnivorous fish, the yellowhead catfish Pelteobagrus fulvidraco, from two connected and five isolated lakes in the flood plain in order to detect the effect of lake fragmentation on the parasite communities. Results: A total of 11 species of helminths were recorded in the stomach and intestine of P. fulvidraco from seven lakes, including two lakes connected with the Yangtze River, i.e. Poyang and Dongting lakes, and five isolated lakes, i.e. Honghu, Liangzi, Tangxun, Niushan and Baoan lakes. Mean helminth individuals and diversity of helminth communities in Honghu and Dongting lakes was lower than in the other five lakes. The nematode Procamallanus fulvidraconis was the dominant species of communities in all the seven lakes. No significant difference in the Shannon-Wiener index was detected between connected lakes (0.48) and isolated lakes (0.50). The similarity of helminth communities between Niushan and Baoan lakes was the highest (0.6708), and the lowest was between Tangxun and Dongting lakes (0.1807). The similarity was low between Dongting and the other lakes, and the similarity decreased with the geographic distance among these lakes. The helminth community in one connected lake, Poyang Lake was clustered with isolated lakes, but the community in Dongting Lake was separated in the tree. Conclusion: The similarity in the helminth communities of this fish in the flood-plain lakes may be attributed to the historical connection of these habitats and to the completion of the life-cycles of this fish as well as the helminth species within the investigated habitats. The diversity and the digenean majority in the helminth communities can be related to the diet of this fish, and to the lacustrine and macrophytic characters of the habitats. The lake isolation from the river had little detectable effect on the helminth communities of the catfish in flood-plain lakes of the Yangtze River. The low similarities in helminth communities between the Dongting Lake and others may just be a reflection of its unique water environment and anthropogenic alterations or fragmentation in this lake.

Plankton Community Succession in Artificial Systems Subjected to Cyanobacterial Blooms Removal using Chitosan-Modified Soils

Using artificial systems to simulate natural lake environments with cyanobacterial blooms, we investigated plankton community succession by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting and morphological method. With this approach, we explored potential ecological effects of a newly developed cyanobacterial blooms removal method using chitosan-modified soils. Results of PCR-DGGE and morphological identification showed that plankton communities in the four test systems were nearly identical at the beginning of the experiment. After applying the newly developed and standard removal methods, there was a shift in community composition, but neither chemical conditions nor plankton succession were significantly affected by the cyanobacteria removal process. The planted Vallisneria natans successfully recovered after cyanobacteria removal, whereas that in the box without removal process did not. Additionally, canonical correspondence analysis indicated that other than for zooplankton abundance, total phosphorus was the most important environmental predictor of planktonic composition. The present study and others suggest that dealing with cyanobacteria removal using chitosan-modified soils can play an important role in controlling cyanobacterial blooms in eutrophicated freshwater systems.

Allelopathic mechanism of pyrogallol to Microcystis aeruginosa PCC7806 (Cyanobacteria): From views of gene expression and antioxidant system

Pyrogallol is a potent allelochemical on Microcystis aeruginosa, but its allelopathic mechanism is not fully known. In order to explore this mechanism, gene expressions for prx, mcyB, psbA, recA, grpE, fabZ under pyrogallol stress were studied, and activities of the main antioxidant enzymes were also measured. The results showed that expression of grpE and recA showed no significant change under pyrogallol stress, while psbA and mcyB were up-regulated at 4 mg L-1. Both prx and fabZ were up-regulated even under exposure to 1 mg L-1 pyrogallol concentration. The activities of superoxide dismutase (SOD) and catalase (CAT) were enhanced under pyrogallol stress. Levels of malodialdehyde (MDA) at 2 and 4 mg L-1 pyrogallol were significantly higher than those of the controls. It was concluded that oxidant damage is an important mechanism for the allelopathic effect of pyrogallol on M. aeruginosa. (c) 2009 Elsevier Ltd. All rights reserved.

Plasma biochemical responses of the planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) to prolonged toxic cyanobacterial blooms in natural waters

The planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) are the attractive candidates for bio-control of plankton communities to eliminate odorous populations of cyanobacteria. However, few studies focused on the health of such fishes in natural water body with vigorous toxic blooms. Blood parameters are useful and sensitive for diagnosis of diseases and monitoring of the physiological status of fish exposed to toxicants. To evaluate the impact of toxic cyanobacterial blooms on the planktivorous fish, 12 serum chemistry variables were investigated in silver carp and bighead carp for 9 months, in a large net cage in Meiliang Bay, a hypereutrophic region of Lake Taihu. The results confirmed adverse effects of cyanobacterial blooms on two phytoplanktivorous fish, which mainly characterized with potential toxicogenomic effects and metabolism disorders in liver, and kidney dysfunction. In addition, cholestasis was intensively implied by distinct elevation of all four related biomarkers (ALP, GGT, DBIL, TBIL) in bighead carp. The combination of LDH, AST activities and DBIL, URIC contents for silver carp, and the combination of ALT. ALP activities and TBIL, DBIL. URIC concentrations for bighead carps were found to most strongly indicate toxic effects from cyanobacterial blooms in such fishes by a multivariate discriminant analysis. (C) 2009 Elsevier B.V. All rights reserved.

Allelopathic interactions between the Potamogeton spp and toxic cyanobacteria (Microcystis aeruginosa)

This study aimed to investigate the allelopathic activities between 3 Potamogeton spp. (Potamogeton maackianus, Potamogeton malaianus and Potamogeton pectinatus) and the toxic cyanobacteria (Microcystis aeruginosa). All Potamogeton spp inhibited the growth of M. aeruginosa in both coexistence and exudates experiments. Inhibition of M. aeruginosa growth by plant exudates depended strongly on the biomass of P malaianus. Initial pH (6.5-9.8) did not influence the inhibitory effects of P. malaianus exudates. However, the M. aeruginosa inhibited the net photosynthesis and respiration of all three pondweed test spp.. The decreases in photosynthesis and respiration were probably caused by the toxic compounds released by M. aeruginosa, rather than its shading effects. The M. aeruginosa also decreased the nutrients (phosphorus and nitrogen) uptake rates of macrophytes. The absorption rates of phosphorus and nitrogen and net photosynthesis were decreased sharply. These results will help to restore submerged plants in eutrophic waters.

Changes in benthic algal communities following construction of a run-of-river dam

Ecological responses to dam construction are poorly understood, especially for downstream benthic algal communities. We examined the responses of benthic algal communities in downstream reaches of a tributary of the Xiangxi River, China, to the construction of a small run-of-river dam. From February 2003 to August 2006, benthic algae, chemical factors, and habitat characteristics were monitored upstream and downstream of the dam site. This period spanned 6 mo before dam construction and 37 mo after dam construction. Benthic algal sampling yielded 199 taxa in 59 genera that belonged to Bacillariophyta, Chlorophyta, and Cyanophyta. Some physical factors (flow velocity, water depth, and channel width) and 3 algal metrics (diatom species richness, Margalef diversity, and % erect individuals) were significantly affected by the dam construction, whereas chemical factors (e.g., NH4-N, total N, SiO2) were not. Nonmetric multidimensional scaling (NMS) ordinations showed that overall algal assemblage structure downstream of the dam sites was similar to that of upstream control sites before dam construction and for 1 year after dam construction (p > 0.05). However, sites belonging to upstream and downstream reaches were well separated on NMS axis 1 during the 2(nd) and 3(rd) years after dam construction. Our results suggest that impacts of dam construction on benthic algal communities took 2 to 3 y to emerge. Further development of a complete set of indicators is needed to address the impact of small-dam construction. Our observations underscore the need for additional studies that quantify ecological responses to dam construction over longer time spans.

Ecology of macrozoobenthic communities in two plateau lakes of Southwest China

Ecological studies on macrozoobenthos were conducted in two small plateau lakes in the Yunnan-Guizhou Plateau, Southwest China: Xingyun Lake (XL), a eutrophic lake whose main source of primary production was phytoplankton (Chl a=99.76 +/- 24.01 mu g/L), and Yangzong Lake (YL), a mesotrophic lake. Sampling was carried out from October 2002 to May 2004. Altogether 23 benthic taxa were identified in XL and 21 taxa in YL. The density of benthos in XL was much lower than that in YL, but the biomass was about equal in the two lakes, being 1 423 ind/m(2) and 8.71 g/m(2) in XL and 4 249 ind/m(2) and 8.60 g/m(2) in YL. The dominant species were Limnodrilus hoffmeisteri, Branchiura sowerbyi, Aulodrilus pluriseta and Chironomus sp. in XL and Limnodrilus hoffmeisteri, Aulodrilus pluriseta and Bellamya sp. in YL. Seasonal fluctuation occurred, showing richer species in summer and winter, but the density and biomass varied in different ways in the two lakes. Analyses on functional feeding groups indicate that collector-gatherers were predominant, but the relative abundances of other groups were different. Stepwise multiple regression analysis demonstrated that the water depth, conductivity and chlorophyll a were the key factors affecting macrozoobenthic abundance in the lakes.

Non-microcystin producing Microcystis wesenbergii (Komarek) Komarek (Cyanobacteria) representing a main waterbloom-forming species in Chinese waters

It is well known that several morphospecies of Microcystis, such as Microcystis aeruginosa (Kutzing) Lemmermann and Microcystis viridis (A. Brown) Lemmermann can produce hepatotoxic microcystins. However, previous studies gave contradictory conclusions about microcystin production of Microcystis wesenbergii (Komarek) Komarek. In the present study, ten Microcystis morphospecies were identified in waterblooms of seven Chinese waterbodies, and Microcystis wesenbergii was shown as the dominant species in these waters. More than 250 single colonies of M. wesenbergii were chosen, under morphological identification, to examine whether M. wesenbergii produce hepatotoxic microcystin by using multiplex PCR for molecular detection of a region (mcyA) of microcystin synthesis genes, and chemical analyses of microcystin content by ELISA and HPLC for 21 isolated strains of M. wesenbergii from these waters were also performed. Both molecular and chemical methods demonstrated that M. wesenbergii from Chinese waters did not produce microcystin. (C) 2007 Elsevier Ltd. All rights reserved.